AbstractThe effect of ethanol upon protein metabolism was investigated using rat hepatocytes purified by centrifugal elutriation and maintained in culture. Hepatocytes maintained in capped culture flasks allowed long‐term studies of the effects of ethanol without cytotoxic damage to cultured cells and the phenotypic integrity of cultured cells was maintained. Ethanol (25–100 mmol/1) markedly inhibited hepatocyte protein synthesis. This inhibition of protein synthesis was not due to defective uptake of amino acids but to alterations to the intracellular redox state. Extracellular redox systems did not regulate hepatocyte protein synthesis. Protein degradation was not affected by ethanol. Glycosaminoglycans secreted into the the medium and extra and intracellular hepatocyte collagen synthesis were markedly stimulated by ethanol; collagen synthesis representing 6–8% of total protein synthesis at high ethanol concentrations. These studies indicate that ethanol exerts selective effects on protein synthesis by hepatocytes in primary culture and suggest that hepatocytes may contribute significantly to perisinusoidal fibrosis in alcohol‐induced liver